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Protective effects of astragaloside IV against amyloid beta1-42 neurotoxicity by inhibiting the mitochondrial permeability transition pore opening.

Sun Q, Jia N, Wang W, Jin H, Xu J, Hu H - PLoS ONE (2014)

Bottom Line: The results showed that pretreatment of AS-IV significantly increased the viability of neuronal cells, reduced apoptosis, decreased the generation of intracellular reactive oxygen species (ROS) and decreased mitochondrial superoxide in the presence of Aβ1-42.Moreover, pretreatment of AS-IV reduced the expression of Bax and cleaved caspase-3 and increased the expression of Bcl-2 in an Aβ1-42 rich environment.These results provide novel insights of AS-IV for the prevention and treatment of neurodegenerative disorders such as AD.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Anatomy and Histo-Embryology, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, China.

ABSTRACT
Mitochondrial dysfunction caused by amyloid β-peptide (Aβ) plays an important role in the pathogenesis of Alzheimer disease (AD). Substantial evidence has indicated that the mitochondrial permeability transition pore (mPTP) opening is involved in Aβ-induced neuronal death and reactive oxygen species (ROS) generation. Astragaloside IV (AS-IV), one of the major active constituents of Astragalus membranaceus, has been reported as an effective anti-oxidant for treating neurodegenerative diseases. However, the molecular mechanisms still need to be clarified. In this study, we investigated whether AS-IV could prevent Aβ1-42-induced neurotoxicity in SK-N-SH cells via inhibiting the mPTP opening. The results showed that pretreatment of AS-IV significantly increased the viability of neuronal cells, reduced apoptosis, decreased the generation of intracellular reactive oxygen species (ROS) and decreased mitochondrial superoxide in the presence of Aβ1-42. In addition, pretreatment of AS-IV inhibited the mPTP opening, rescued mitochondrial membrane potential (ΔΨm), enhanced ATP generation, improved the activity of cytochrome c oxidase and blocked cytochrome c release from mitochondria in Aβ1-42 rich milieu. Moreover, pretreatment of AS-IV reduced the expression of Bax and cleaved caspase-3 and increased the expression of Bcl-2 in an Aβ1-42 rich environment. These data indicate that AS-IV prevents Aβ1-42-induced SK-N-SH cell apoptosis via inhibiting the mPTP opening and ROS generation. These results provide novel insights of AS-IV for the prevention and treatment of neurodegenerative disorders such as AD.

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AS-IV attenuates Aβ1-42-induced increase of ROS generation in SK-N-SH cells.A. Confocal fluorescence images of intracellular ROS stained with DCFH-DA. Mitochondria were counterstained with Mitotracker Red (red fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. B. Quantification analysis for DCFH-DA fluorescence intensity. #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). C. Confocal fluorescence images of mitochondrial superoxide stained with MitoSOX Red. Mitochondria were counterstained with Mitotracker Green (green fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µ µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. D. Quantification analysis for MitoSOX Red fluorescence intensity #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). Scale bar = 10 µm.
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pone-0098866-g007: AS-IV attenuates Aβ1-42-induced increase of ROS generation in SK-N-SH cells.A. Confocal fluorescence images of intracellular ROS stained with DCFH-DA. Mitochondria were counterstained with Mitotracker Red (red fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. B. Quantification analysis for DCFH-DA fluorescence intensity. #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). C. Confocal fluorescence images of mitochondrial superoxide stained with MitoSOX Red. Mitochondria were counterstained with Mitotracker Green (green fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µ µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. D. Quantification analysis for MitoSOX Red fluorescence intensity #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). Scale bar = 10 µm.

Mentions: To analyze whether AS-IV attenuates cell death by blocking ROS generation, we examined the level of intracellular ROS by using H2DCF-DA fluorescent dye. The lowest ROS level was detected from the vehicle group. Compared with the vehicle group, the Aβ1-42 group showed significantly higher level of green fluorescence intensity (P<0.01). In the presence of AS-IV, DCF intensity was significantly decreased in a dose-dependent manner compared with the Aβ1-42 group (P<0.01). However, pretreatment of AS-IV did not completely reverse the ROS level compared with that in the vehicle group (P<0.01) (Fig. 7A, B). 50 µM AS-IV alone treatment did not show an insult to SK-N-SH cells.


Protective effects of astragaloside IV against amyloid beta1-42 neurotoxicity by inhibiting the mitochondrial permeability transition pore opening.

Sun Q, Jia N, Wang W, Jin H, Xu J, Hu H - PLoS ONE (2014)

AS-IV attenuates Aβ1-42-induced increase of ROS generation in SK-N-SH cells.A. Confocal fluorescence images of intracellular ROS stained with DCFH-DA. Mitochondria were counterstained with Mitotracker Red (red fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. B. Quantification analysis for DCFH-DA fluorescence intensity. #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). C. Confocal fluorescence images of mitochondrial superoxide stained with MitoSOX Red. Mitochondria were counterstained with Mitotracker Green (green fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µ µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. D. Quantification analysis for MitoSOX Red fluorescence intensity #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). Scale bar = 10 µm.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
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pone-0098866-g007: AS-IV attenuates Aβ1-42-induced increase of ROS generation in SK-N-SH cells.A. Confocal fluorescence images of intracellular ROS stained with DCFH-DA. Mitochondria were counterstained with Mitotracker Red (red fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. B. Quantification analysis for DCFH-DA fluorescence intensity. #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). C. Confocal fluorescence images of mitochondrial superoxide stained with MitoSOX Red. Mitochondria were counterstained with Mitotracker Green (green fluorescence). (a) vehicle; (b) 50 µM AS-IV; (c) 5 µ µM Aβ1-42; (d, e, f) 10, 25, 50 µM AS-IV+ 5 µM Aβ1-42, respectively. D. Quantification analysis for MitoSOX Red fluorescence intensity #P<0.01 vs vehicle; *P<0.01 vs Aβ1-42 (n = 8). Scale bar = 10 µm.
Mentions: To analyze whether AS-IV attenuates cell death by blocking ROS generation, we examined the level of intracellular ROS by using H2DCF-DA fluorescent dye. The lowest ROS level was detected from the vehicle group. Compared with the vehicle group, the Aβ1-42 group showed significantly higher level of green fluorescence intensity (P<0.01). In the presence of AS-IV, DCF intensity was significantly decreased in a dose-dependent manner compared with the Aβ1-42 group (P<0.01). However, pretreatment of AS-IV did not completely reverse the ROS level compared with that in the vehicle group (P<0.01) (Fig. 7A, B). 50 µM AS-IV alone treatment did not show an insult to SK-N-SH cells.

Bottom Line: The results showed that pretreatment of AS-IV significantly increased the viability of neuronal cells, reduced apoptosis, decreased the generation of intracellular reactive oxygen species (ROS) and decreased mitochondrial superoxide in the presence of Aβ1-42.Moreover, pretreatment of AS-IV reduced the expression of Bax and cleaved caspase-3 and increased the expression of Bcl-2 in an Aβ1-42 rich environment.These results provide novel insights of AS-IV for the prevention and treatment of neurodegenerative disorders such as AD.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Anatomy and Histo-Embryology, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, China.

ABSTRACT
Mitochondrial dysfunction caused by amyloid β-peptide (Aβ) plays an important role in the pathogenesis of Alzheimer disease (AD). Substantial evidence has indicated that the mitochondrial permeability transition pore (mPTP) opening is involved in Aβ-induced neuronal death and reactive oxygen species (ROS) generation. Astragaloside IV (AS-IV), one of the major active constituents of Astragalus membranaceus, has been reported as an effective anti-oxidant for treating neurodegenerative diseases. However, the molecular mechanisms still need to be clarified. In this study, we investigated whether AS-IV could prevent Aβ1-42-induced neurotoxicity in SK-N-SH cells via inhibiting the mPTP opening. The results showed that pretreatment of AS-IV significantly increased the viability of neuronal cells, reduced apoptosis, decreased the generation of intracellular reactive oxygen species (ROS) and decreased mitochondrial superoxide in the presence of Aβ1-42. In addition, pretreatment of AS-IV inhibited the mPTP opening, rescued mitochondrial membrane potential (ΔΨm), enhanced ATP generation, improved the activity of cytochrome c oxidase and blocked cytochrome c release from mitochondria in Aβ1-42 rich milieu. Moreover, pretreatment of AS-IV reduced the expression of Bax and cleaved caspase-3 and increased the expression of Bcl-2 in an Aβ1-42 rich environment. These data indicate that AS-IV prevents Aβ1-42-induced SK-N-SH cell apoptosis via inhibiting the mPTP opening and ROS generation. These results provide novel insights of AS-IV for the prevention and treatment of neurodegenerative disorders such as AD.

Show MeSH
Related in: MedlinePlus